T Cell Modulation of Renal Ischemia Reperfusion Injury

描述(由申请人提供)：肾缺血再灌注损伤(IRI)是导致自体和同种移植肾急性肾功能衰竭(ARF)的主要原因。我们和其他人已经证明了T细胞在实验性肾ARF中的重要调节作用。下一阶段是了解T细胞在肾脏IRI中的作用机制。我们假设T细胞通过T细胞受体(TCR)-主要组织相容性复合体(MHC)的抗原特异性相互作用来调节肾脏IRI，并产生与其他特定淋巴细胞协同工作的炎症介质。具体目标1：我们假设TCR-MHC复合体通过抗原特异性反应介导T细胞参与肾脏IRI。我们已有初步数据表明，TCR受体和MHC II类分子直接影响肾脏IRI的预后。我们将使用已建立的肾脏IRI小鼠模型和具有特定基因敲除的小鼠，并评估肾脏、分子和细胞的反应。我们还将进行T细胞过继移植研究，并使用骨髓嵌合体。我们将使用CD4TCR转基因小鼠来探讨抗原特异性和T细胞激活在IRI中的作用。具体目标2：我们假设早期T细胞的运输、激活和细胞因子的产生介导了肾脏IRI后的后续损伤。我们将使用我们新开发的淋巴细胞分离技术来评估T细胞进入缺血后肾脏的情况。我们将比较T细胞与其他白细胞的转运，并从蛋白和mRNA水平表征体外肾脏T细胞的激活状态和促炎细胞因子的产生。过继转移技术将用于直接测试这些T细胞细胞因子的功能作用。具体目标3：我们假设T细胞与NKT和B细胞相互作用，以充分表达IRI后的肾脏损伤。我们将使用NKT和B细胞功能缺陷的小鼠，并进行过继转移研究。我们将使用以前限制这些研究的新开发工具，在野生型小鼠身上进行激动剂、阻滞剂和耗竭剂的补充实验。这是一个新的方向，将导致对启动缺血后肾脏炎症的非常早期的细胞步骤的新见解。长期目标将是确定促进肾脏IRI损伤过程的抗原和分子途径，并开发针对ARF的靶向免疫疗法。

DESCRIPTION (provided by applicant): Kidney ischemia reperfusion injury (IRI) is a major cause of acute renal failure (ARF) in both native and allograft kidney. We and others have demonstrated an important modulatory role for T cells in experimental kidney ARF. The next stage is to understand the mechanisms underlying the role for T cells in renal IRI. We hypothesize that T cells modulate renal IRI by trafficking into ischemic kidney, becoming activated through antigen specific interactions through the T cell receptor (TCR)-major histocompatibility complex (MHC), and produce inflammatory mediators that work in concert with other specific lymphocytes. Specific Aim 1: We hypothesize that the TCR-MHC complex mediates T cell participation in renal IRI through antigen specific responses. We have preliminary data that TCR receptors, as well as MHC class II directly influence outcome of renal IRI. We will use an established mouse model of renal IRI and mice with specific knockouts, and evaluate renal, molecular and cellular responses. We will also perform T cell adoptive transfer studies and use bone marrow chimeras. We will use CD4 TCR transgenic mice to probe the role of antigen specificity and T cell activation in IRI. Specific Aim 2: We hypothesize that early T cell trafficking, activation and production of cytokines mediates subsequent injury following renal IRI. We will use our new developed lymphocyte isolation technique to evaluate trafficking of T cells into postischemic kidney. We will compare T cell trafficking with that of other leukocytes, and characterize the renal T cells ex vivo in terms of activation status and production of proinflammatory cytokines at the protein and mRNA level. Adoptive transfer techniques will be used to directly test the functional role of these T cell cytokines. Specific Aim 3: We hypothesize that T cells interact with NKT and B cells for full expression of renal injury after IRI. We will use mice deficient in NKT and B cell function, and perform adoptive transfer studies. We will perform complementary experiments with agonists, blockers and depleting agents in wild type mice using new developed tools that previously limited these studies. This is a novel direction that will lead to new insights into the very early cellular steps that initiate postischemic kidney inflammation. The long term goal will be to identify the antigens and molecular pathways that fuel the injury processes in renal IRI, and develop targeted immune therapies for ARF.